Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complex
Aim:This study evaluated the use of polymerase chain reaction for cryptococcal meningitis diagnosis in clinical samples.Materials and methods:The sensitivity and specificity of the methodology were evaluated using eight Cryptococcus neoformans/C. gattiispecies complex reference strains and 165 cere- brospinal fluid samples from patients with neurological diseases divided into two groups: 96 patients with cryptococcal meningitis and AIDS; and 69 patients with other neurological opportunistic diseases (CRL/AIDS). Two primer sets were tested (CN4-CN5 and the multiplex CNa70S-CNa70A/CNb49S-CNb-49A that amplify a specific product for C. neoformansand another for C. gattii).Results:CN4-CN5 primer set was positive in all Cryptococcusstandard strains and in 94.8% in DNA samples from cryptococcal meningitis and AIDS group. With the multiplex, no 448-bp product of C. gattiiwas observed in the clinical samples of either group. The 695 bp products of C. neoformanswere observed only in 64.6% of the cryptococcal meningitis and AIDS group. This primer set was negative for two standard strains. The specificity based on the negative samples from the CTL/AIDS group was 98.5% in both primer sets.Conclusions:These data suggest that the CN4/CN5 primer set was highly sensitive for the identification of C. neoformans/C. gattiispecies complex in cerebrospinal fluid samples from patients with clinical suspicion of cryptococcal meningitis.
| Main Authors: | , , , , , |
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| Format: | Digital revista |
| Language: | English |
| Published: |
Brazilian Society of Infectious Diseases
2015
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| Online Access: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702015000100062 |
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| Summary: | Aim:This study evaluated the use of polymerase chain reaction for cryptococcal meningitis diagnosis in clinical samples.Materials and methods:The sensitivity and specificity of the methodology were evaluated using eight Cryptococcus neoformans/C. gattiispecies complex reference strains and 165 cere- brospinal fluid samples from patients with neurological diseases divided into two groups: 96 patients with cryptococcal meningitis and AIDS; and 69 patients with other neurological opportunistic diseases (CRL/AIDS). Two primer sets were tested (CN4-CN5 and the multiplex CNa70S-CNa70A/CNb49S-CNb-49A that amplify a specific product for C. neoformansand another for C. gattii).Results:CN4-CN5 primer set was positive in all Cryptococcusstandard strains and in 94.8% in DNA samples from cryptococcal meningitis and AIDS group. With the multiplex, no 448-bp product of C. gattiiwas observed in the clinical samples of either group. The 695 bp products of C. neoformanswere observed only in 64.6% of the cryptococcal meningitis and AIDS group. This primer set was negative for two standard strains. The specificity based on the negative samples from the CTL/AIDS group was 98.5% in both primer sets.Conclusions:These data suggest that the CN4/CN5 primer set was highly sensitive for the identification of C. neoformans/C. gattiispecies complex in cerebrospinal fluid samples from patients with clinical suspicion of cryptococcal meningitis. |
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